Cited by 2,892Carcinogens and Genotoxicity AssessmentDNA Repair MechanismsEffects of Radiation Exposure

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The Comet Assay for DNA Damage and Repair: Principles, Applications, and Limitations

Collins, Andrew R.·2004·Molecular Biotechnology

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Abstract

The comet assay (single-cell gel electrophoresis) is a simple method for measuring deoxyribonucleic acid (DNA) strand breaks in eukaryotic cells. Cells embedded in agarose on a microscope slide are lysed with detergent and high salt to form nucleoids containing supercoiled loops of DNA linked to the nuclear matrix. Electrophoresis at high pH results in structures resembling comets, observed by fluorescence microscopy; the intensity of the comet tail relative to the head reflects the number of DNA breaks. The likely basis for this is that loops containing a break lose their supercoiling and become free to extend toward the anode. The assay has applications in testing novel chemicals for genotoxicity, monitoring environmental contamination with genotoxins, human biomonitoring and molecular epidemiology, and fundamental research in DNA damage and repair. The sensitivity and specificity of the assay are greatly enhanced if the nucleoids are incubated with bacterial repair endonucleases that recognize specific kinds of damage in the DNA and convert lesions to DNA breaks, increasing the amount of DNA in the comet tail. DNA repair can be monitored by incubating cells after treatment with damaging agent and measuring the damage remaining at intervals. Alternatively, the repair activity in a cell extract can be measured by incubating it with nucleoids containing specific damage.

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[36]36. Novotná, B. (2000) Increased DNA fragmentation detected by comet assay in peripheral blood of heterozygotes for a frameshift mutation in the DNA repair protein nibrin. DNA Repair Workshop Smolenice, Slovakia, October 2000, Abstract p. 33.

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Authors: Collins, A. R.
Affiliations: University of Oslo (NO)
Corresponding author: Collins, Andrew R.
ORCID: Collins, Andrew R.
Venue: Molecular Biotechnology
Publisher: Springer Science and Business Media LLC
Published: 2004
Volume: 26
Issue: 3
Pages: 249-261
ISSN: 1073-6085 (ISSN-L)
DOI: 10.1385/mb:26:3:249
OpenAlex: W2034485370
Cited by: 2,892
References: 46
Citations by year: 2026: 91 · 2025: 104 · 2024: 119 · 2023: 134 · 2022: 138 · 2021: 157 · 2020: 146 · 2019: 168 · 2018: 164 · 2017: 174 · 2016: 161 · 2015: 179 · 2014: 156 · 2013: 162 · 2012: 171
FWCI: 27.05
Type: journal article
Language: en
Open access: closed
Locations: Molecular Biotechnology (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
Keywords: comet assay, DNA strand breaks, DNA damage, DNA repair, genotoxicity testing, genotoxins, human biomonitoring, molecular epidemiology, DNA supercoiling
MeSH terms: Animals, DNA, DNA Damage, DNA Mutational Analysis, DNA Repair, Environmental Monitoring, Environmental Pollutants, Humans, Mutagens, Apoptosis, Molecular Epidemiology, Comet Assay

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The Comet Assay for DNA Damage and Repair: Principles, Applications, and Limitations

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